Sterilizing immunity against SARS-CoV-2 in hamsters conferred by a novel recombinant subunit vaccine

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   ScreenIT

   Dec 21, 2020

   SciScore for 10.1101/2020.12.18.423552: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained. Consent: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained.
   Randomization	Cynomolgus monkey vaccination: Eight Cynomolgus monkeys were allocated randomly into four groups (one female and one male per group).
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Cynomolgus monkey vaccination: Eight Cynomolgus monkeys were allocated …

   SciScore for 10.1101/2020.12.18.423552: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained. Consent: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained.
   Randomization	Cynomolgus monkey vaccination: Eight Cynomolgus monkeys were allocated randomly into four groups (one female and one male per group).
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Cynomolgus monkey vaccination: Eight Cynomolgus monkeys were allocated randomly into four groups (one female and one male per group).
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   For Tfh cells analysis, the following antibodies were used: rat anti-mouse CXCR5 (BD, 551961), biotin-conjugated anti-rat IgG2a (Biolegend, 407504), APC conjugated streptavidin (eBioscience, 17-4317-82), FITC conjugated anti-mouse CD4 (Biolegend, 100406), PE/cy7 conjugated anti-mouse CD8α (Biolegend, 100722), PerCP/cy5.5 conjugated anti-mouse TCRβ (Biolegend, 109228), BV421 conjugated anti-mouse PD-1 (Biolegend, 135221) and LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit.	anti-mouse CXCR5 suggested: (BioLegend Cat# 145511, RRID:AB_2562127) anti-rat IgG2a suggested: (BioLegend Cat# 202529, RRID:AB_10899572) anti-mouse CD4 suggested: (BioLegend Cat# 100406, RRID:AB_312691) anti-mouse CD8α suggested: (Bio X Cell Cat# BE0117, RRID:AB_10950145) anti-mouse TCRβ suggested: None anti-mouse PD-1 suggested: (BioLegend Cat# 135221, RRID:AB_2562568)
   The binding activity of polyclonal antibodies in serum was defined as the dilution fold required to achieve 50% of the maximal signal (ED50).	ED50 suggested: None
   Pseudovirus neutralizing antibody titer measurement: Vesicular stomatitis virus (VSV) based SARS-CoV-2 pseudotyping-particles (VSVpp) were produced according to our previous study (49).	VSV suggested: None
   Serum samples were collected weekly for biochemical and antibody analyses, including measurement of anti-spike IgG, pseudovirus neutralizing antibody, and receptor-blocking antibody titers during the Week 1 to 10.	anti-spike IgG suggested: None
   Immunohistochemical staining was performed by using the mouse monoclonal anti-SARS-CoV-2 N proteins antibody.	anti-SARS-CoV-2 N suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Subsequently, the culture supernatants of CHO cells were subjected to purification by Ni Sepharose Excel column (Cytiva).	CHO suggested: CLS Cat# 603479/p746_CHO, RRID:CVCL_0213)
   The BHK21-hACE2 cells were used to perform the pseudovirus neutralization assay.	BHK21-hACE2 suggested: None
   Briefly, the hACE2-mRuby3 293T (293T-ACE2iRb3) cells were seeded in poly-D-lysine pretreated CellCarrier-96 Black plate at 2×104 cells per well overnight.	293T suggested: RRID:CVCL_YZ65)
   After removing half of the medium from the pre-seeded 293T-ACE2iRb3 cells, pipette 50 μl of the mixture into the wells and incubate 1 hour at 37 °C containing 5% CO2.	293T-ACE2iRb3 suggested: None
   Briefly, Vero cells were seeded in a 6-well plate at 105 cells per well.	Vero suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Mouse immunization: For antibody response evaluation, BALB/c mice were maintained in a specific pathogen-free (SPF) environment and immunized with various proteins at 1 μg/dose with Al001 or FH002C through intramuscular injection, following an immunization schedule of one priming dose at Week 0 plus two boosters at Weeks 2 and 4.	BALB/c suggested: None
   To germinal center response characterization, the T follicular helper cells (Tfh), germinal center B cells (GCB), and plasma cells in LNs from immunized C57BL/6 mice at 1-week after single immunization were analyzed by Flurescence Activated Cell Sorting (FACS).	C57BL/6 suggested: None
   Software and Algorithms
   Sentences	Resources
   Subsequently, cells were fixed and permeated by using Fixation/Permeabilization Solution Kit (BD Biosciences), and further stained with BV421-conjugated anti-mouse IL-4 (Biolegend, 504127) and, PE-conjugated anti-mouse IFN-γ (Biolegend, 505826).	BD Biosciences suggested: (BD Biosciences, RRID:SCR_013311)
   Finally, the samples were measured by BD LSRFortessa X-20 Flow Cytometer (BD), and the data were analyzed by FlowJo V10.6.0.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Statistical analyses were conducted in GraphPad Prism 8 software.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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   Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

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   Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   Results from rtransparent:

   • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
   • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
   • No protocol registration statement was detected.

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   Version published to 10.1101/2020.12.18.423552 on bioRxiv

   Dec 20, 2020